Year: 2023
Type: dissertation
Abstract: Living cells can be engineered into cell-based therapeutics by utilizing synthetic genetic elements and protein effectors that reprogram cell activity to be able to treat diseases. Modular design of different components in these engineered systems creates a toolkit that enables efficient customization and scalability to different applications. This thesis describes the creation of a synthetic promoter, SFCp, that has reduced de novo synthesis costs, a short length and a constitutive expression profile achieved by compression of transcription factor motifs associated with high constitutive expression. Next, we describe the application of this promoter to cancer immunotherapy using single-chain interleukin-12. The anti-cancer ability of stably transduced tumour cells with SFCp-expressed interleukin-12 was assessed in two mouse models, N2A neuroblastoma and 4T1 metastatic breast cancer, demonstrating the modularity and potential for SFCp to be used in engineered in vivo applications. Utilizing syngeneic tumour cells as a delivery strategy for immunotherapeutics addresses the four cell-based therapeutic key pillars: therapeutic effect, targetability, immune-cloaking, and kill-switch. The development of low-cost synthetic genetic elements expands the modular components available to facilitate future engineered system designs, with compelling application to cell-based therapeutics that are capable of much more complex function and regulation than simple molecular drugs.
Source: TSpace (University of Toronto)
Author: Lam, Chee Ka Candice
Institution: University of Toronto
Language: English
Subfield: Biotechnology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7132997352